A multiplex polymerase chain reaction to detect and differentiate Campylobacter fetus subspecies fetus and Campylobacter fetus -species venerealis: use on UK isolates of C. fetus and other Campylobacter spp.

Willoughby, Kim; Pf, Nettleton; Mm, Quirie; Ma, Maley; Foster, Geoffrey; Toszeghy, M.; Dg, Newell

doi:10.1111/j.1365-2672.2005.02680.x

Cited by 40 publications

(59 citation statements)

References 19 publications

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“…venerealis and C. fetus subsp. fetus are plasmid encoded (25,26), although more recent studies have questioned this proposal (42). This MLST scheme indicates that the differences between the two subspecies are not merely plasmid encoded but are also reflected in the allelic profiles (STs) of the housekeeping genes.…”

Section: Discussionmentioning

confidence: 99%

Clonal Nature of Campylobacter fetus as Defined by Multilocus Sequence Typing

Bergen¹,

et al. 2005

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Campylobacter fetus can be divided into the subspecies C. fetus subsp. fetus and C. fetus subsp. venerealis. C. fetus subsp. fetus causes sporadic infections in humans and abortion in cattle and sheep and has been isolated from a variety of sites in different hosts. C. fetus subsp. venerealis is host restricted, being isolated mainly from the genital tracts of cattle, and is the causative agent of bovine genital campylobacteriosis. Despite differences in niche preference, microbiological subspecies differentiation has proven difficult. Different typing methods divided C. fetus isolates into different subgroups, depending on the methods used. The relative value of these methods can be assessed by the evolutionary relationship of isolates belonging to the genus; therefore, we developed a multilocus sequence typing (MLST) scheme for C. fetus. This scheme was applied to 140 C. fetus isolates previously typed by amplified fragment length polymorphism (AFLP) analysis. A total of 14 different sequence types (STs) were identified, and these exhibited low levels of inter-ST genetic diversity, with only 22 variable sites in 3,312 nucleotides. These MLST data indicate that C. fetus is genetically homogeneous compared to the homogeneity of other Campylobacter species. The two C. fetus subspecies were extremely closely related genetically, but ST-4 was associated only with C. fetus subsp. venerealis, which represents a "bovine" clone. The C. fetus subsp. fetus isolates studied were more diverse in terms of their STs, and the STs correlated with epidemiological relationships. Congruence was observed among C. fetus subspecies, sap type, and ST;

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Section: Discussionmentioning

confidence: 99%

Clonal Nature of Campylobacter fetus as Defined by Multilocus Sequence Typing

Bergen¹,

et al. 2005

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“…venerealis from C. fetus subsp. fetus is extremely difficult (72). A single phenotypic test is recommended by the OIE for typing the subspecies (66); thus, international efforts currently focus on optimizing genotypic tests.…”

Section: Discussionmentioning

confidence: 99%

Interbacterial Macromolecular Transfer by the Campylobacter fetus subsp. venerealis Type IV Secretion System

et al. 2011

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We report here the first demonstration of intra-and interspecies conjugative plasmid DNA transfer for Campylobacter fetus. Gene regions carried by a Campylobacter coli plasmid were identified that are sufficient for conjugative mobilization to Escherichia coli and C. fetus recipients. A broader functional range is predicted. Efficient DNA transfer involves the virB9 and virD4 genes of the type IV bacterial secretion system encoded by a pathogenicity island of C. fetus subsp. venerealis. Complementation of these phenotypes from expression constructions based on the promoter of the C. fetus surface antigen protein (sap) locus was temperature dependent, and a temperature regulation of the sap promoter was subsequently confirmed under laboratory conditions. Gene transfer was sensitive to surface or entry exclusion functions in potential recipient cells carrying IncP␣ plasmid RP4 implying functional relatedness to C. fetus proteins. The virB/virD4 locus is also known to be involved in bacterial invasion and killing of cultured human cells in vitro. Whether specifically secreted effector proteins contribute to host colonization and infection activities is currently unknown. Two putative effector proteins carrying an FIC domain conserved in a few bacterial type III and type IV secreted proteins of pathogens were analyzed for secretion by the C. fetus or heterologous conjugative systems. No evidence for interbacterial translocation of the Fic proteins was found.

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“…Under these stringent conditions, the results of the glycine tolerance test and PCR were compared, and there was a complete correspondence between the results of both methods. Some groups (5,13,16,19) used a multiplex PCR assay to identify and differentiate the C. fetus isolates. In our experience, problems can occur because of the insufficient optimization of the assay, such as by the use of different primer concentrations.…”

Section: Discussionmentioning

confidence: 99%

Identification of Campylobacter fetus Subspecies by Phenotypic Differentiation and PCR

Schulze

Bagon²,

Müller

et al. 2006

J Clin Microbiol

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The species Campylobacter fetus is divided into the subspecies C. fetus subsp. venerealis and C. fetus subsp. fetus, which differ in their epidemiologies and clinical importance. The differences between these subspecies make accurate distinction between the two essential. First, the value of seven key tests for the traditional differentiation of C. fetus was investigated. Afterwards, the results of the phenotypic differentiation and PCR were compared to address the question of the reliability of this PCR assay. Altogether, 103 C. fetus isolates were investigated, including the type strains of C. fetus subsp. fetus and C. fetus subsp. venerealis. Depending on the result of the glycine tolerance test, the isolates could be separated into 81 C. fetus subsp. venerealis isolates (glycine intolerant) and 22 C. fetus subsp. fetus isolates (glycine tolerant). For all C. fetus subsp. venerealis strains tested, the results of the selenite reduction assay and sensitivity to metronidazole and cefoperazone completely agreed with the results of the glycine tolerance test (correspondence, 100%). Seventy-three C. fetus subsp. venerealis isolates did not grow at 42°C (correspondence, 90.1%), but eight isolates showed a faintly discernible, flat, dark gray growth. For 22 C. fetus subsp. fetus isolates, the results of additional phenotypic tests only partly agreed with the results of the glycine tolerance test. For C. fetus subsp. fetus the results of the glycine tolerance test showed a relatively good correspondence with those of the selenite reduction assay (correspondence, 81.8%), assays for cefoperazone resistance (correspondence, 86.4%), and assays for growth at 42°C (correspondence, 81.8%). The results of the glycine tolerance test and PCR completely agreed for the 103 C. fetus isolates tested. We conclude that at present the traditional phenotypic characterization of C. fetus subspecies under strongly defined conditions remains indispensable, but this PCR assay constitutes a valuable adjunctive technique for the confirmation of phenotypic results.

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A multiplex polymerase chain reaction to detect and differentiate Campylobacter fetus subspecies fetus and Campylobacter fetus -species venerealis: use on UK isolates of C. fetus and other Campylobacter spp.

Cited by 40 publications

References 19 publications

Clonal Nature of Campylobacter fetus as Defined by Multilocus Sequence Typing

Clonal Nature of Campylobacter fetus as Defined by Multilocus Sequence Typing

Interbacterial Macromolecular Transfer by the Campylobacter fetus subsp. venerealis Type IV Secretion System

Identification of Campylobacter fetus Subspecies by Phenotypic Differentiation and PCR

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